CN111789257A - Morchella polysaccharide extract, effervescent tablets, and preparation methods and applications of morchella polysaccharide extract and effervescent tablets - Google Patents
Morchella polysaccharide extract, effervescent tablets, and preparation methods and applications of morchella polysaccharide extract and effervescent tablets Download PDFInfo
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- CN111789257A CN111789257A CN202010771751.9A CN202010771751A CN111789257A CN 111789257 A CN111789257 A CN 111789257A CN 202010771751 A CN202010771751 A CN 202010771751A CN 111789257 A CN111789257 A CN 111789257A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/40—Effervescence-generating compositions
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Sustainable Development (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
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- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a morchella polysaccharide extract, an effervescent tablet, and preparation methods and applications thereof, and belongs to the field of functional foods. The preparation method of the morchella polysaccharide extract comprises the following steps: extracting Morchella esculenta at low temperature, precipitating with ethanol, decolorizing, removing protein, purifying, and separating to obtain Morchella esculenta polysaccharide extract. The morchella polysaccharide effervescent tablet is prepared by the steps of crushing, granulating, drying and granulating, totally mixing and tabletting the morchella polysaccharide extract and tablet auxiliary materials. The morchella polysaccharide extract obtained by the invention has high purity and bioactivity, and the extraction yield and the extraction efficiency are higher than those of the traditional extraction method, and the energy is saved compared with the traditional extraction method. The morchella esculenta is separated and purified to prepare the functional effervescent tablet, and the functional effervescent tablet is used for carrying out systematic experimental researches on kidney tonifying, yang strengthening and the like, discussing the pharmacological effects of the functional effervescent tablet, and has a certain reference value on the application of morchella esculenta polysaccharide in the health care functions of kidney tonifying and yang strengthening.
Description
Technical Field
The invention relates to the field of functional foods, and particularly relates to a morchella polysaccharide extract, an effervescent tablet, and preparation methods and applications thereof.
Background
Morchella rare fungi used for both food and medicine has been recorded in compendium of materia Medica of Li Shizhen for a long time, and is named Morchella due to its uneven surface like Morchella. The morchella has high nutritional and medicinal values, according to determination, the morchella contains various nutritional components such as polysaccharide, protein, crude fat, amino acid and the like, the morchella polysaccharide is a main active component of the morchella, and is internationally called as a health food, so that the morchella has the effects of benefiting intestines and stomach, promoting digestion, reducing phlegm, regulating qi, tonifying kidney, strengthening yang, tonifying brain, refreshing and the like after being frequently eaten, and also can build the body, prevent cold and enhance the immunity of the human body. Therefore, the method has great development and application prospects in the fields of food, medicine, health care, cosmetics and the like. The extraction of the morchella is relatively similar to that of other edible fungi, and the research is not many, and the traditional high-temperature single-tank extraction is more.
The effervescent tablet is a novel tablet in China, the reaction of organic acid and alkali type carbonate (hydrogen) salt is used as an effervescent disintegrant, the effervescent reaction is immediately carried out after the effervescent tablet is placed in water, and a large amount of carbon dioxide gas is generated and released. In addition, the effervescent tablet needs strong moisture absorption resistance, otherwise the effervescent effect can be influenced, and auxiliary materials with strong moisture absorption resistance and good compressibility have great research space.
The traditional Chinese medicine considers that the kidney is the root of the congenital foundation and the life, and has extremely important position and function in the human physiology. This is mainly because the mutual dependence of yin-yang, water, fire, original qi and essence and blood in the kidney struggle against each other to promote the whole life activity of human body. In the Nei Jing, it is mentioned that the kidney, mainly water, is hidden by the essence of the five zang-organs and six fu-organs; the essence of yin bestows on the blood of life and bestows on the body, so it is expensive. The experimental study of the morchella on the functions of tonifying kidney and strengthening yang is not reported in detail temporarily.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provide a preparation method of a morchella polysaccharide extract.
The invention also aims to provide the morchella polysaccharide extract obtained by the preparation method and application thereof.
The invention further aims to provide the effervescent tablet containing the morchella polysaccharide extract, which has good taste and convenient carrying, and has the functions of tonifying kidney and strengthening yang.
The purpose of the invention is realized by the following technical scheme:
a preparation method of morchella polysaccharide extract comprises the following steps: extracting Morchella esculenta at low temperature, precipitating with ethanol, decolorizing, removing protein, purifying, and separating to obtain Morchella esculenta polysaccharide extract; the method specifically comprises the following steps:
(1) carrying out ultrasonic-assisted low-temperature continuous extraction on the morchella esculenta powder to obtain a morchella esculenta polysaccharide aqueous extract;
(2) concentrating the morchella polysaccharide water extract obtained in the step (1), precipitating with ethanol, and collecting precipitate to obtain a morchella polysaccharide crude extract;
(3) dissolving the morchella polysaccharide crude extract obtained in the step (2) to obtain a morchella polysaccharide dissolving solution; decolorizing the crude morchella polysaccharide dissolved solution, and carrying out solid-liquid separation to obtain a decolorized crude morchella polysaccharide solution;
(4) carrying out enzymolysis deproteinization on the decolored morchella crude polysaccharide solution obtained in the step (3) by using protease, and filtering to obtain a decolored deproteinized crude polysaccharide solution;
(5) and (4) concentrating the decolored deproteinized crude polysaccharide solution obtained in the step (4), and drying to obtain the morchella polysaccharide, namely the morchella polysaccharide extract.
The morchella esculenta powder in the step (1) is prepared by the following steps: drying and pulverizing Morchella esculenta fruiting body, making into coarse powder, and sieving to obtain Morchella esculenta powder.
The drying temperature is 40-60 ℃; preferably 50 deg.c.
The screening is preferably performed by a 10-40-mesh screen; more preferably a 20 mesh screen.
The ultrasonic-assisted low-temperature continuous extraction method in the step (1) comprises the following steps: the three extraction tanks are mutually connected through vacuum pump pipes, are named as an extraction tank A, an extraction tank B and an extraction tank C respectively, and contain ultrasonic equipment; placing morchella powder materials in three extraction tanks respectively; adding water into the extraction tank A to perform first extraction of the extraction tank A, and feeding the obtained extract into a storage tank; adding water into the extraction tank A to perform secondary extraction, taking the obtained extract as an extraction solvent, feeding the extract into an extraction tank B, performing primary extraction in the extraction tank B, and feeding the obtained extract into a storage tank; adding water into the extraction tank A to perform third extraction of the extraction tank A, feeding the obtained extract into the extraction tank B to perform second extraction of the extraction tank B, feeding the obtained extract into the extraction tank C to perform first extraction of the extraction tank C, and feeding the obtained extract into a storage tank; discharging residue in tank A, and adding Morchella esculenta again; adding water into the extraction tank B to perform third extraction of the extraction tank B, feeding the obtained extract into the extraction tank C to perform second extraction of the extraction tank C, feeding the obtained extract into the extraction tank A to perform first extraction of the extraction tank A (second tank raw material), and feeding the obtained extract into a storage tank; discharging slag in the tank B, and adding the morchella raw material again; adding water into the extraction tank C for the third extraction of the extraction tank C, feeding the obtained extract into the extraction tank A for the second extraction of the extraction tank A (second tank raw material), feeding the obtained extract into the extraction tank B as an extraction solvent for the first extraction of the extraction tank B, and feeding the obtained extract into a storage tank; discharging slag in a tank C, and adding the morchella raw material again; the extraction is sequentially circulated, and the morchella raw material added into each tank is extracted for three times.
The extraction conditions are preferably as follows: carrying out ultrasonic-assisted extraction in the water bath time of the extraction tank at 50-75 ℃ for 0.5-3 h; preferably a water bath at 65 ℃ for 1 h.
The power of the ultrasonic equipment is 200-600 w.
The power of the ultrasonic waves extracted for the first time is preferably 400-600 w; more preferably 500 w.
The power of the second extraction is preferably 300-400 w; more preferably 350 w.
The power of the third extraction is preferably 200-300 w; more preferably 250 w.
The first extraction, the second extraction and the third extraction refer to the first extraction, the second extraction and the third extraction in the extraction tank A, B, C.
The ultrasonic-assisted extraction time is 10-30 min for each extraction; preferably, each extraction is sonicated for 20 min.
The ratio of the first extraction material to the first extraction material in the tank A is preferably 1: 8-10.
The ratio of the second extraction material to the liquid in the tank A is preferably 1: 10-12.
The ratio of the material to the liquid extracted from the tank A for the third time is preferably 1: 12-15.
The ratio of the material to the liquid extracted in the tank B for the third time is preferably 1: 10-15; more preferably 12 to 15.
The ratio of the material to the liquid extracted in the third time of the tank C is preferably 1: 8-15; more preferably 12 to 15.
The total dosage of water in the morchella polysaccharide aqueous extract in the step (1) is calculated according to the ratio of the total dosage of morchella powder to the total dosage of water being 3: 40-50 (g: mL); preferably, the ratio of Morchella esculenta powder to water is 3: 44 (g: mL).
The concentration in step (2) is preferably vacuum concentration.
The vacuum concentration conditions are preferably as follows: the vacuum degree is-0.06-0.10 MPa, and the temperature is 50-70 ℃; more preferably-0.08 MPa, 65 ℃.
The concentration degree in the step (2) is preferably concentrated to a thick extract with the density of 1.18-1.28 g/mL; more preferably, the concentration is carried out until the density is 1.20 to 1.23 g/mL.
The alcohol in the alcohol precipitation in the step (2) is preferably ethanol; preferably a 95% (v/v) ethanol solution.
The dosage of the ethanol is preferably calculated according to the volume percentage of the ethanol in an ethanol precipitation system of 70-85 percent; preferably, the alcohol is 75% by volume of the alcohol in the alcohol precipitation system.
The time for alcohol precipitation in the step (2) is preferably 8-14 h; more preferably 10 h.
The manner of collecting the precipitate in the step (2) is preferably centrifugation; the centrifugation conditions are as follows: centrifuging at 2000-5000 rpm for 10-20 min; more preferably, centrifugation is carried out at 4000rpm for 15 min.
The dissolved solvent in step (3) is preferably water; more preferably purified water.
In the morchella crude polysaccharide dissolving solution in the step (3), calculating a morchella crude polysaccharide extract and a solvent according to a volume ratio of 1: 4-6; preferably at a volume ratio of 1: 5.
And (3) decoloring in the step (3) by using macroporous resin.
The macroporous resin is preferably food grade D315 macroporous resin.
The dosage of the macroporous resin is calculated according to the volume of 1/3-1/2 of the morchella crude polysaccharide dissolving solution.
The decolorization is preferably carried out under stirring.
The decolorizing condition is preferably 200-400 rpm stirring for 2-4 h; more preferably 300rpm for 3 hours.
The protease in step (4) preferably comprises at least one of trypsin, papain and bromelain; more preferably papain.
When the protease is trypsin, adjusting the pH value to 6-8; preferably, the pH is adjusted to 7.0.
When the protease is papain, adjusting the pH value to 4-6; preferably, the pH is adjusted to 5.5.
When the protease is bromelain, adjusting the pH value to 6-8; preferably, the pH is adjusted to 6.5.
The dosage of the trypsin is 10-25U/mg; preferably 15U/mg.
The dosage of the papain is 10-25U/mg; preferably 20U/mg.
The using amount of the bromelain is 10-25U/mg; preferably 20U/mg.
The temperature of the enzymolysis deproteinization in the step (4) is preferably 50-65 ℃; more preferably 55 deg.c.
The time for enzymolysis deproteinization in the step (4) is preferably 1-3 h; more preferably 2 h.
The concentration in step (5) is preferably vacuum concentration.
The vacuum concentration conditions are preferably as follows: the vacuum degree is-0.06 to-0.10 MPa, and the temperature is 50 to 70 ℃; more preferably-0.08 MPa, 65 ℃.
The drying in step (5) is preferably freeze-drying.
A Morchella polysaccharide extract is prepared by the above preparation method.
The morchella polysaccharide extract is applied to the preparation of a health-care functional preparation for tonifying kidney and strengthening yang.
A health promotion functional preparation for invigorating kidney and tonifying yang comprises adjuvants and the above Morchella polysaccharide extract.
The auxiliary materials are used for assisting in preparing the morchella polysaccharide extract into different dosages, preferably including but not limited to tablets.
The tablet is preferably an effervescent tablet.
An effervescent tablet containing Morchella polysaccharide extract comprises component A, sweetener and lubricant; the component A comprises the following components in parts by mass: 220-300 parts of morchella esculenta polysaccharide extract, 260-320 parts of citric acid, 160-200 parts of sodium bicarbonate, 180-240 parts of cane sugar and 200-290 parts of isomaltitol; the content of the sweetening agent is 0-1.0% of the mass of the component A, and the content of the lubricating agent is 0-0.7% of the mass of the component A.
The sweetener is preferably sucralose.
The content of the sweetening agent is preferably 0.5-1.0 percent of the mass of the component A; more preferably 0.7% by mass of the A component.
The lubricant is preferably magnesium stearate.
The content of the lubricant is preferably 0.2 to 0.7 percent of the mass of the component A; more preferably 0.5% by mass of the A component.
The preparation method of the effervescent tablet containing the morchella polysaccharide extract comprises the following steps:
mixing the ground and sieved sweetener, citric acid, cane sugar and isomalt, and adding a wetting agent to prepare wet granules A; mixing morchella polysaccharide and sodium bicarbonate, adding a wetting agent, and preparing wet granules B;
(II) drying and finishing the wet granules A and the wet granules B obtained in the step (I) respectively to obtain two kinds of dry granules;
(III) mixing the two dry particles obtained in step (II) with a lubricant to obtain a total mixture;
and (IV) tabletting the total mixture obtained in the step (III) to obtain the effervescent tablet containing the morchella polysaccharide extract.
The wetting agent is preferably a 50% (v/v) ethanol solution.
The dosage of the wetting agent is preferably calculated according to the ratio of the wetting agent to the required wetting material being 0.2-0.3 mL: 1 g; more preferably, the ratio of wetting agent to required wetting material is 0.25mL to 1 g.
The citric acid and the cane sugar in the step (I) need to be dried before being crushed and sieved.
The drying temperature is 55-60 ℃.
The degree of drying is as follows: drying citric acid and sucrose to water content below 2%.
The sieving in the step (I) is to sieve the mixture through a sieve of 60-100 meshes; preferably through an 80 mesh screen.
The mixing equipment described in step (I) is preferably a high-speed mixing wet granulator.
The adjusting pressure of the high-speed mixing wet granulator is 0.4 Mpa.
The rotating speed of a stirring paddle of the high-speed mixing wet granulator is 150-250 rpm; preferably 210 rpm.
The chopping rotating speed of the high-speed mixing wet granulator is 1000-1600 rpm; preferably 1400 rpm.
And (3) drying equipment in the step (II) is a cabinet type drying box.
The drying temperature in the step (II) is 50-65 ℃; preferably 60 deg.c.
The degree of drying in step (ii) is preferably: drying until the moisture content is not higher than 3%.
The whole granulation apparatus described in step (II) is preferably a rocking granulator.
The mesh size of the whole granules in step (ii) is preferably 20 mesh.
The mixing apparatus described in step (III) is preferably a three-dimensional mixer.
The mixing conditions in step (III) are preferably: mixing for 15-25 min at the main shaft rotating speed of 24 r/min; more preferably a spindle speed of 24r/min for 20 min.
The tabletting equipment in the step (IV) is preferably a full-automatic tabletting machine.
The tablet weight of the tablet compressed in the step (IV) is preferably 2.4 g/tablet.
Compared with the prior art, the invention has the following advantages and effects:
1. the morchella polysaccharide extract is obtained by low-temperature extraction, alcohol precipitation, decoloration, deproteinization, purification and separation of morchella, and the obtained morchella polysaccharide extract has high purity and biological activity, and has higher extraction yield and extraction efficiency than the traditional extraction method, and is more energy-saving than the traditional extraction method.
2. The morchella polysaccharide effervescent tablet is prepared by crushing, granulating, finishing, totally mixing and tabletting the morchella polysaccharide extract and tablet auxiliary materials. The morchella polysaccharide effervescent tablet is novel in dosage form, products taking morchella as a raw material on the market have no dosage form, the preparation technology is simple, the process is stable, and the morchella polysaccharide effervescent tablet is suitable for large-scale production.
3. In the preparation process, isomaltitol is used as a main forming agent to replace commonly used better tablet auxiliary materials such as D-mannitol, lactose, part of cane sugar and the like, and the extract raw material with high moisture absorption can effectively prevent moisture absorption and agglomeration in the tabletting process, and the pressed tablets have moderate hardness and good wear resistance and are free from sticking during tabletting. And the sugar content is reduced, and the product is healthier and safer.
4. The prepared morchella polysaccharide extract effervescent tablet has good taste, is convenient to carry, is simple to eat, has high safety of raw and auxiliary materials, and can be taken for a long time.
5. The morchella esculenta is separated and purified to prepare the functional effervescent tablet, and the functional effervescent tablet is used for carrying out systematic experimental researches on kidney tonifying, yang strengthening and the like, discussing the pharmacological effects of the functional effervescent tablet, and has a certain reference value on the application of morchella esculenta polysaccharide in the health care functions of kidney tonifying and yang strengthening.
Drawings
FIG. 1 is a graph showing the deproteinization effect of various deproteinizing enzymes on a decolorized morchella crude polysaccharide solution.
FIG. 2 is a graph showing the effect of Morchella polysaccharide extracts prepared by different extraction methods on the elimination of hydroxyl radicals.
FIG. 3 is a graph showing the effect of Morchella polysaccharide extracts prepared by different extraction methods on the elimination of DPPH free radicals.
Fig. 4 is a graph of the absorption kinetics of different effervescent tablets containing morchella polysaccharide extract.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Example 1 preparation of Morchella polysaccharide extract
(1) Selecting wild morchella sporocarp in Yuxi City in Yunnan province, drying at 50 ℃ to constant weight, crushing into coarse powder, and sieving with a 20-mesh sieve to obtain morchella granules with uniform size.
(2) Weighing 1 part by mass of morchella esculenta powder respectively, placing the morchella esculenta powder in A, B, C tanks (A, B, C tanks each have extraction conditions of water bath at 65 ℃ for 1h), and connecting A, B, C extraction tanks with each other through vacuum pump pipes, wherein the extraction tanks contain ultrasonic devices (A, B, C tanks have power of 500w, 350w and 250w for the first extraction, the second extraction and the third extraction respectively, and the time of ultrasonic treatment is 20min at the beginning of each extraction). Adding the morchella raw material into each tank, extracting for three times, feeding the extract obtained by the first extraction of the morchella raw material into a storage tank, and taking the extract obtained by the second extraction and the third extraction as an extraction solvent of the next tank to perform circular extraction according to the principle. The method comprises the following specific steps:
1) the ultrasonic-assisted low-temperature extraction is characterized in that in a morchella polysaccharide aqueous extract, the ratio of total dosage of morchella powder to total dosage of water is 3: 40.
A. Extracting in tank A at a ratio of 1: 8 (g: mL, extraction solvent is purified water; the same applies below) for the first time, and storing the extractive solution in a storage tank.
B. Performing the first round of second extraction in tank A at a ratio of 1: 10 (g: mL, extraction solvent is purified water), introducing the extractive solution into tank B, performing the first round of first extraction in tank B, and introducing the extractive solution into storage tank.
C. Adding water into tank A at a ratio of 1: 12 for the first round of the third extraction, introducing the extractive solution into tank B for the first round of the second extraction, introducing the extractive solution into tank C for the first round of the first extraction, and introducing the final extractive solution into storage tank.
D. And discharging slag in the tank A, and adding 1 part by mass of morchella raw material again.
E. Adding water into tank B at a ratio of 1: 12 for the first round of third extraction, adding the extractive solution into tank C for the first round of second extraction, adding the obtained extractive solution into tank A for the first round of first extraction (second tank of Morchella esculenta raw material), and adding the obtained extractive solution into storage tank.
F. And discharging slag in the tank B, and adding 1 part by mass of morchella raw material again.
G. Adding water into tank C at a ratio of 1: 12, performing first round of third extraction, performing second round of second extraction, taking the obtained extractive solution as extraction solvent, performing second round of first extraction, and storing in storage tank.
H. And C, discharging slag in the tank C, and adding the morchella raw material again.
I. Adding water into tank A at a ratio of 1: 12 for the third extraction of the second round, feeding the obtained extractive solution into tank B, performing the second extraction of the second round, feeding the obtained extractive solution into tank C, performing the first extraction of the second round, and feeding the obtained extractive solution into storage tank.
J. Discharging residue in tank A, and adding Morchella esculenta again.
K. Adding water at a ratio of 1: 12 into tank B for second round of third extraction, adding the extractive solution into tank C for second round of second extraction, adding the obtained extractive solution into tank A for third round of first extraction (third tank of Morchella esculenta raw material), and adding the obtained extractive solution into storage tank.
L, B, discharging slag, and adding 1 part by mass of morchella raw material again.
And M, adding water into the tank C at a material-liquid ratio of 1: 12 for a second round of third extraction, feeding the obtained extracting solution into the tank A for a third round of second extraction, feeding the obtained extracting solution serving as an extraction solvent into the tank B for a third round of first extraction in the tank B, and feeding the obtained extracting solution into a storage tank.
N, C discharging residue, and adding Morchella esculenta.
And O, adding water into the tank A at a material-liquid ratio of 1: 12 for third extraction in a third round, feeding the obtained extracting solution into the tank B, carrying out second extraction in the third round in the tank B, feeding the obtained extracting solution into the tank C, carrying out first extraction in the third round in the tank C, and feeding the obtained extracting solution into a storage tank.
And P, adding water into the tank B at a ratio of material to liquid of 1: 10 for third extraction, adding the extracting solution into the tank C for second extraction, and adding the obtained extracting solution into a storage tank.
And Q, adding water into the tank C at a ratio of 1: 8 for third extraction, and feeding the obtained extract into a storage tank.
The embodiment is a three-wheel cycle, the cycle is about to be ended at the step Q, and the amount of added water is properly reduced; if the circulation is continued, the circulation is performed according to the steps C-O.
2) The ultrasonic-assisted low-temperature extraction is characterized in that in a morchella polysaccharide aqueous extract, the ratio of total dosage of morchella powder to total dosage of water is 3: 44.
The steps are basically the same as 1) group, the difference is only that the adding amount of water is different, and the using amount of the water in different steps is as follows:
A(1︰9)、B(1︰11)、C(1︰13)、E(1︰13)、G(1︰13)、I(1︰13)、K(1︰13)、M(1︰13)、O(1︰13)、P(1︰11)、Q(1︰9)。
3) the ultrasonic-assisted low-temperature extraction is characterized in that in a morchella polysaccharide aqueous extract, the ratio of total dosage of morchella powder to total dosage of water is 3: 50.
The steps are basically the same as 1) group, the difference is only that the adding amount of water is different, and the using amount of the water in different steps is as follows:
A(1︰10)、B(1︰12)、C(1︰15)、E(1︰15)、G(1︰15)、I(1︰15)、K(1︰15)、M(1︰15)、O(1︰15)、P(1︰12)、Q(1︰10)。
(3) and (3) carrying out vacuum concentration on the morchella aqueous extract obtained in the step (2) until the density is 1.20-1.23 g/mL, the vacuum degree is-0.08 MPa, and the temperature is 65 ℃. Adding 95% (v/v) ethanol solution into the concentrated solution to make ethanol concentration reach 75% (v/v), standing for 10h, centrifuging at 4000rpm for 15min, and collecting precipitate to obtain polysaccharide crude extract.
(4) Dissolving the morchella polysaccharide crude extract obtained in the step (3) with purified water with the volume 5 times that of the morchella polysaccharide crude extract to obtain a morchella crude polysaccharide dissolving solution, adding 1/3 food-grade D315 macroporous resin with the volume of the morchella crude polysaccharide dissolving solution, stirring by using a magnetic stirrer (300rpm, 3 hours), filtering, and performing solid-liquid separation to obtain a decolored morchella crude polysaccharide solution.
(5) Dividing the decolorized morchella crude polysaccharide solution obtained in the step (4) into three groups, respectively adding trypsin (pH value of 7.0 and enzyme dosage of 15U/mg), papain (pH value of 5.5 and enzyme dosage of 20U/mg) and bromelain (pH value of 6.5 and enzyme dosage of 20U/mg), deproteinizing for 2h at 55 ℃, and then respectively determining the polysaccharide loss rate and the protein removal rate by a phenol-sulfuric acid method and a Coomassie brilliant blue G-250 method. The results of the determination after enzymolysis with trypsin, papain and bromelain, respectively, are shown in FIG. 1. The results show that: the 3 proteases have good deproteinizing effect, wherein, the deproteinization of the papain is more efficient and has less loss, namely, the effect is better.
(6) And (4) carrying out vacuum concentration on the dehydrated deproteinized polysaccharide solution obtained in the step (5) under the conditions of-0.08 MPa and 65 ℃ to obtain a thick extract, and then carrying out freeze drying to obtain the morchella polysaccharide. Measuring polysaccharide content and polysaccharide yield, calculating solvent usage amount, time consumption and energy consumption, and comparing the indexes of morchella polysaccharide prepared by a traditional high-temperature extraction method; wherein the traditional method for extracting Morchella esculenta polysaccharide at high temperature comprises steaming and decocting, and three times of intermittent extraction, wherein the material-to-liquid ratio of the three times of extraction is 1: 10, 1: 8, and 1: 6 respectively. The traditional high-temperature extraction method of morchella polysaccharide described in this embodiment is a cooking method (i.e., the traditional high-temperature extraction in table 1), wherein: the cooking temperature is 100 ℃, the time is 23h, the feed-liquid ratio of the morchella esculenta powder to the water is 1: 24, and the results are shown in table 1.
The results of the above-mentioned index measurements are shown in Table 1. Wherein:
TABLE 1
The results show that: compared with the traditional high-temperature extraction method, the method has the advantages that the extraction yield is high, the polysaccharide content is high, the consumed time is short, and the energy consumption is reduced by adopting an ultrasonic-assisted low-temperature extraction (namely low-temperature continuous extraction) method.
Example 2 Morchella polysaccharide antioxidant assay
The morchella polysaccharide obtained in example 1 by traditional high-temperature extraction and ultrasonic-assisted low-temperature extraction at a material-liquid ratio of 3: 44 is prepared into solutions (sample solutions) with a series of concentrations of 0.2mg/ml, 0.6mg/ml, 0.8mg/ml and 1.0 mg/ml.
(1) Hydroxyl radical scavenging ability
Refer to the measurement method in Leonian et al, 7 studies on the in vitro antioxidant activity of hot water extracts of Ganoderma from different sources, and slightly improve the method. Accurately sucking 1mL of sample solution with different concentrations, and respectively adding 1mL of FeSO with concentration of 6mmol/L4Solution and 1mL of 6mmol/L H2O2Shaking the solution, standing for 10min, adding 1mL of 6mmol/L salicylic acid solution, shaking, standing for 30min, replacing salicylic acid with distilled water in the control group, replacing sample with distilled water in the blank group, and measuring absorbance at 510nm with ultraviolet spectrophotometer. Calculating the clearance rate of the hydroxyl free radical by a calculation formula:
wherein: a. theBlank spaceBlank absorbance (distilled water instead of sample solution);
Asample (I)Is the absorbance value of the added sample liquid;
AcontrolAbsorbance values for the control group.
(2) DPPH radical scavenging ability
Reference is made to Ma et al "Optimization for the extraction of polysaccharides from Garoderma lucidum and the anti-oxidant and anti-viral activities" with minor modifications. Sucking 2mL of polysaccharide solution, adding 2mL of DPPH solution with the concentration of 0.2mmol, mixing uniformly, reacting in the dark for 30min, and measuring the absorbance at 517 nm. The control group was prepared by replacing DPPH with absolute ethanol, and the blank group was prepared by replacing polysaccharide solution with distilled water. DPPH free radical clearance calculation formula:
Ablank spaceBlank absorbance (distilled water instead of sample solution);
Asample (I)Is the absorbance value of the added sample liquid;
AcontrolAbsorbance values for the control group.
The hydroxyl radical elimination rate and DPPH radical elimination rate of the extracts obtained by the low-temperature continuous extraction and the traditional high-temperature extraction under different concentrations are respectively shown in fig. 2 and fig. 3. The results show that the extract obtained by the low-temperature continuous extraction in example 1 has higher antioxidant capacity than the extract obtained by the traditional high-temperature extraction of the control group.
Example 3 preparation of effervescent tablet containing Morchella polysaccharide extract
(1) The effervescent tablets containing the morchella polysaccharide extract are prepared by adding the raw materials according to the mass parts in the table 2.
TABLE 2 (units are parts by mass)
The preparation method comprises the following steps: mixing crushed and sieved (sieved by a 80-mesh sieve) citric acid (the moisture content is lower than 2%), cane sugar (the moisture content is lower than 2%), isomaltitol and sucralose in a high-speed wet granulator (the pressure is adjusted to be 0.4Mpa, the rotating speed of a stirring paddle is 210rpm, and the chopping speed is 1400rpm), adding a wetting agent (50% (v/v) ethanol solution), and preparing wet granules according to the amount of the wetting agent (mL) and the mixture (g) of the citric acid, the cane sugar, the isomaltitol and the sucralose. Mixing Morchella esculenta polysaccharide and sodium bicarbonate high-speed wet granulator (adjusting pressure to 0.4Mpa, stirring paddle rotation speed to 210rpm, cutting speed to 1400rpm), adding wetting agent (50% (v/v) ethanol solution), and making into wet granule according to wetting agent (mL): sodium bicarbonate + Morchella esculenta polysaccharide mixture (g): 0.25: 1. Drying the two groups of wet granules in cabinet type drying oven respectively (drying at 60 deg.C until water content is not higher than 3%), and granulating with swing granulator (mesh number is 20). Pouring the two groups of dry granules and lubricant magnesium stearate into a three-dimensional mixer to mix (mixing for 20min at the main shaft rotating speed of 24 r/min) to obtain a total mixture, tabletting by using a full-automatic tabletting machine, wherein the tablet weight is 2.4 g/tablet to obtain effervescent tablets containing the morchella polysaccharide extract, and the tablet weight can also be adjusted according to needs.
The effervescent tablets 1-3 and the reference 1-3 contain morchella polysaccharide extract, and the performance test is as follows:
1. determination of moisture absorption
Reference is made to the research methods of Yufrin and the like in the influence of lactose and mannitol on the hygroscopicity of the Gushuling granules. Precisely weighing about 2g of a sample to be inspected, evenly spreading the sample in a weighing bottle which is dried to constant mass, opening a cover, placing the bottle in a dryer for more than 12 hours to achieve dehumidification balance, and reserving the bottle for later use. At 25 ℃, the supersaturated NaCl solution was placed at the bottom of the dryer for 48 hours to bring the interior thereof to a constant relative humidity of 75%. And precisely weighing the weighing bottle with the sample, putting the weighing bottle into the dryer (the weighing bottle cap is opened), taking out the weighing bottle after 4h, 8h, 12h, 24h, 48h and 72h, tightly covering the weighing bottle, and precisely weighing the weighing bottle and the sample. An average of 3 replicates of each sample was taken and the percent moisture absorption calculated. The time is plotted on the abscissa and the moisture absorption percentage is plotted on the ordinate, and the moisture absorption kinetics curve is shown in FIG. 4.
The percent moisture absorption was calculated as follows: the moisture absorption percentage is ═ mass after moisture absorption-mass before moisture absorption)/mass before moisture absorption ] × 100%.
2. Friability test
According to the fourth part of the Chinese pharmacopoeia, the friability test method of tablets is generally known. Taking 10 effervescent tablets, blowing off powder falling off from the tablets by using a blower, precisely weighing, placing in a cylinder of a tablet four-purpose tester (Jinan Xin Beixi Biotechnology Co., Ltd.), and rotating for 4 min. Taking out, removing powder by the same method, precisely weighing, reducing weight loss not to exceed 1%, and not detecting fracture, crack and crushed sheet. This test was generally carried out only 1 time. If the weight loss exceeds 1%, the weight loss should be measured again 2 times, and the average weight loss of 3 times should not exceed 1%, and no fracture, crack or crushed piece should be detected.
3. Hardness test
According to the fourth part of the Chinese pharmacopoeia, the general rule is "tablet hardness test method". The tablet is radially fixed between two cross bars for measuring the hardness of a tablet four-purpose tester (Jinan Xin Beixi Biotechnology Co., Ltd.), a movable column bar in the tablet four-purpose tester radially pressurizes the tablet in the horizontal direction by virtue of a spring, when the tablet is broken, the spring of the movable bar stops pressurizing, and the pressure indicated by a dial of the tester is the hardness of the tablet. The number of 3 tablets was measured and averaged. Hardness requirements for effervescent tablet storage and transport: the higher the hardness, the better the effervescence is acceptable. The results of the preparation and testing are shown in Table 3.
TABLE 3
| Sample (I) | Whether sticking or not | Friability (%) | Hardness (N) |
| |
Without sticking | 0.39 | 44.78 |
| Effervescent tablet 2 | Without sticking | 0.41 | 42.65 |
| |
Without sticking | 0.42 | 40.67 |
| |
Slight sticking | 0.83 | 32.69 |
| Control 2 | Slight sticking | 0.78 | 29.58 |
| |
Sticking punch | 1.04 | 20.57 |
From the above results, it can be seen that: the effervescent tablets 1-3 added with isomalt have good moisture absorption resistance, the moisture absorption resistance of the granules is slightly better than that of a control combination of sucrose and D-mannitol or lactose substituted isomalt which are single auxiliary materials, and the effervescent tablets are more favorable for storage and effervescent foamability of products; the formula combination of the effervescent tablets 1-3 does not stick to impact in the preparation process, the extruded tablets have higher hardness, are not easy to crack, have low friability and are not easy to fall off, so the compressibility of the effervescent tablets 1-3 added with isomalt is superior to that of a reference combination.
Product indexes such as sensory, effervescence state and mouthfeel of drink after effervescence of the effervescent tablets 1-3 are evaluated, and the results are shown in table 4:
TABLE 4 effervescent tablet product index evaluation
Example 4 Kidney-tonifying and Yang-strengthening function test
An experiment on the functions of invigorating the kidney and strengthening yang was carried out by using the effervescent tablet 2 containing the morchella polysaccharide extract prepared in example 3.
(one) sample dosage
The normal daily dose for an adult is 2.4g (1 tablet) and is 60 kg, the dosage design is shown in Table 5.
Table 5: mouse sample administration dose equivalent converted according to body weight
KM mice, male and female, purchased at the experimental animals center of university of zhongshan.
SD rats, male, purchased at the experimental animals center of university of zhongshan.
Guilu Bushen Wan (Tortoise and deer Kidney tonifying pill), Guangzhou city Huacheng pharmaceutical factory, national medicine Standard Z44020148.
(II) test of influence of test sample on mouse capture rate
Selecting 40 male mice and 40 female mice of healthy KM mice; the 40 male mice were randomly grouped into blank control group, low, medium and high dose groups, 10 mice in each group, and the male mice were in solitary state during the whole experiment. The experimental group was given test samples at the dosages shown in table 5; purified water was administered once daily to the blank control group for 10 days by continuous gavage. After the last administration for 45min, a female mouse (estrus after vaginal smear detection) is placed in each cage of the independent male mouse, and then observed for 1h, the capturing latency and capturing frequency of the female mouse are recorded, the capturing rate of each group is calculated, the results are statistically processed, and the differences among the groups are compared.
(III) test of influence of test sample on mouse conception
Selecting 40 male mice and 40 female mice of healthy KM mice; the 40 male mice were randomly grouped into blank control group, low, medium and high dose groups, 10 mice in each group, and the male mice were in solitary state during the whole experiment. The experimental group was given test samples at the dosages shown in table 5; purified water was administered once daily to the blank control group for 10 days by continuous gavage. After the last administration, 1 male mouse and 1 female mouse with reproductive capacity (estrus period by vaginal smear) are combined in a cage, whether vaginal suppository exists in the female mouse is checked on the 2 nd day after the combination, the female mouse can be detected as a pregnant mouse by vaginal smear microscopic examination, and the pregnant mouse is recorded; the number of pregnant mice was recorded for 5 consecutive days, the results were statistically processed and the differences between groups were compared.
(IV) test of penis erection by test sample
Selecting 54 healthy SD male rats, wherein 10 of the SD male rats are used as a sham operation group (a normal control group), not removing testicles at both sides, only opening the skin and then sewing, carrying out intraperitoneal injection anesthesia on the rest male rats by using 10% chloral hydrate normal saline, disinfecting scrotum skin by using 75% alcohol, removing testicles at both sides, randomly dividing the male rats into 5 groups, carrying out low, medium and high dose groups on samples, carrying out model control group and positive drug guilu kidney-tonifying pill control group, carrying out 4 rats on the guilu kidney-tonifying pill control group, and carrying out 10 rats in the rest groups. Rats were injected with 2 ten thousand units/kg of penicillin intramuscularly after surgery and treated with antibiotics for 3 days.
Each group of animals was orally administered the test drug by gavage every day, the sham-operated group and the model control group were administered purified water of equal volume, and the remaining groups of rats were administered the doses shown in Table 5. After 30 days of drug administration, penis electrical stimulation experiments are carried out on the rats of each group, electrodes of the JL-B type electrical stimulator are placed at the penis parts of the rats, and stimulation parameters are as follows: voltage 50V, frequency 30Hz, wave width 0.2ms, amplitude 1, local electrical stimulation was given and the erection time from the start of stimulation to the penis, i.e. the erection latency, was recorded. After the experiment is finished, the fast is carried out for 12 hours, eyes are picked off to take blood, and serum is prepared for detecting the Zn content. Immediately after sacrifice, the penis was taken to prepare a homogenate for total nitric oxide synthase assay of the penis tissue. Prostate ventral lobes, seminal vesicle glands, levator ani muscles were isolated, weighed and the correlation index calculated. Penile erection latency and correlation index were statistically processed and compared for differences between groups.
Example results:
(I) test results of the Capture Rate of test sample to mouse
The capture latency of high-dose mice in the sample is obviously shortened, and compared with a blank control group in the same period, the high-dose mice have significant differences (P <0.05 and P < 0.001); meanwhile, the middle and high dose groups of the samples can increase the capture frequency of the female mice within 1h, and have significant difference (P <0.01 and P <0.001) compared with the blank control group (see Table 6). The above suggests that the test sample has a certain effect of promoting yang-strengthening.
Table 6: results of the test sample-to-mouse capture rate experiment: (
n＝10)
Note: p <0.01, p <0.001 compared to the blank control group (t-test).
(II) test result of mouse conception test by test sample
After one estrus female mouse was placed in each group of solitary mice daily overnight, the number of pregnant female mice in each dosing group was significantly increased (see table 7). In the sample, the total pregnancy number and the pregnancy rate of the high-dose female mice have significant difference compared with those of a blank control group at the same period (P <0.01 and P < 0.001). The test sample is prompted to have a certain promotion effect on strengthening yang.
Table 7: results of the test sample-to-mouse capture rate experiment: (
n＝10)
Note: p compared to blank control group<0.01，***p<0.001(X2Test).
(III) test result of penis erection by test sample to rat
Except for the sham operation group, the rats in other groups were subjected to testicular removal operation, and after the operation, the rats in all groups recovered well, and the results of measuring the penile erection time, i.e., the penile erection latency, after 30 days of administration (see table 8), the results of measuring NOS by serum zinc and penis homogenate (see table 9), and the results of weighing the sacrificed sex organs of the rats and calculating the visceral index (see table 10); wherein, the serum zinc is measured by flame atomic absorption spectrophotometry, and the wavelength is 213.9 nm; penile NOS total nitric oxide synthase NOS activity was determined using penile tissue homogenates.
Table 8: experimental results of test samples on penile erection latency in rats: (
n＝10)
Note: p <0.05, p <0.01, p <0.001 compared to model controls (t-test).
As shown in Table 8, the penis erection latency of the model control group rats is significantly prolonged, and compared with the same-phase normal control group, the difference is significant (P <0.001), which indicates that the sexual function of the rats is significantly inhibited after the testicles are extirpated. The penis erection latent period of the rat can be obviously shortened by the medium dosage and the high dosage of the sample, and compared with a contemporary model control group, the penis erection latent period has significant differences (P <0.05, P <0.01 and P <0.001), which indicates that the two samples have certain regulating effect on the sexual function of the rat. The positive medicine Guilu Bushen Wan can obviously shorten the erection latent period of the penis of a rat, has obvious difference (P is less than 0.001) compared with a contemporary model control group, and prompts that the Guilu Bushen Wan has certain yang-tonifying effect and accords with the clinical effect.
Table 9: test results of NOS assay of test sample on rat serum zinc and penis homogenate: (
n＝10)
Note: p <0.05, p <0.01 compared to model control group (t-test).
As can be seen from Table 9, the serum zinc levels of the rats in the normal control group and the rats in the model control group are similar, and have no significant difference (P >0.05), which suggests that the rats have limited influence on the serum zinc ion level within one month after the testis is removed. The serum zinc levels of both test samples were either high or low, but there was no significant difference (P >0.05) compared to the contemporary model control group, which is an individual difference of the animals. The result of total nitric oxide synthase NOS activity measurement by penis tissue homogenate shows that the NOS level of a normal control group of rats is increased and has a significant difference (P <0.05) compared with that of a contemporary model control group, which indicates that the activity of NOS is influenced after the kidney yang deficiency of the rats caused by testis removal. The high dosage of the sample can obviously improve the activity of NOS, and compared with a contemporary model control group, the sample has significant differences (P <0.05 and P <0.01), which indicates that the sample has a certain regulating effect on the sexual function of castrated rats. The positive medicine Guilu Bushen Wan can obviously improve the activity of penis NOS, has significant difference (P <0.05) compared with a contemporary model control group, and prompts that the Guilu Bushen Wan has a certain yang-tonifying effect and conforms to the clinical effect.
Table 10: the result of the test sample on the index of the organ index of the sex organ of the rat with kidney-yang deficiency caused by testis removal (
n＝10)
P <0.05, p <0.01 compared to model control group (t-test).
The experimental results in table 10 show that the rats in the model control group have atrophy of prostate ventral lobe, seminal vesicle gland, levator ani muscle and glandula preputiales in different degrees, and have significant differences (P <0.05 and P <0.01) compared with the rats in the same normal control group; after the testis is removed, the level of the male hormone of the rat is obviously reduced, and the sexual organ is atrophied. After 30 days of administration, the sex organ index of rats can be obviously increased by the medium dose and the high dose of the sample, and compared with a contemporary model control group, the sex organ index of rats has significant differences (P <0.05 and P <0.01), which indicates that the samples have certain hormone-like effects on castrated rats. The positive medicine Guilu Bushen Wan has higher indexes of prostate, abdominal lobe, levator ani and glandular preputiales, has significant difference (P <0.05) compared with a contemporary model control group, and prompts that the Guilu Bushen Wan has a certain hormone-like effect and accords with the clinical effect thereof.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A preparation method of a morchella polysaccharide extract is characterized by comprising the following steps: extracting Morchella esculenta at low temperature, precipitating with ethanol, decolorizing, removing protein, purifying, and separating to obtain Morchella esculenta polysaccharide extract; the method specifically comprises the following steps:
(1) carrying out ultrasonic-assisted low-temperature continuous extraction on the morchella esculenta powder to obtain a morchella esculenta polysaccharide aqueous extract;
(2) concentrating the morchella polysaccharide water extract obtained in the step (1), precipitating with ethanol, and collecting precipitate to obtain a morchella polysaccharide crude extract;
(3) dissolving the morchella polysaccharide crude extract obtained in the step (2) to obtain a morchella polysaccharide dissolving solution; decolorizing the crude morchella polysaccharide dissolved solution, and carrying out solid-liquid separation to obtain a decolorized crude morchella polysaccharide solution;
(4) carrying out enzymolysis deproteinization on the decolored morchella crude polysaccharide solution obtained in the step (3) by using protease, and filtering to obtain a decolored deproteinized crude polysaccharide solution;
(5) and (4) concentrating the decolored deproteinized crude polysaccharide solution obtained in the step (4), and drying to obtain the morchella polysaccharide, namely the morchella polysaccharide extract.
2. The method of claim 1, wherein the step (1) of ultrasonic-assisted low-temperature continuous extraction comprises: the three extraction tanks are mutually connected through vacuum pump pipes, are named as an extraction tank A, an extraction tank B and an extraction tank C respectively, and contain ultrasonic equipment; placing morchella powder materials in three extraction tanks respectively; adding water into the extraction tank A to perform first extraction of the extraction tank A, and feeding the obtained extract into a storage tank; adding water into the extraction tank A to perform secondary extraction, taking the obtained extract as an extraction solvent, feeding the extract into an extraction tank B, performing primary extraction in the extraction tank B, and feeding the obtained extract into a storage tank; adding water into the extraction tank A to perform third extraction of the extraction tank A, feeding the obtained extract into the extraction tank B to perform second extraction of the extraction tank B, feeding the obtained extract into the extraction tank C to perform first extraction of the extraction tank C, and feeding the obtained extract into a storage tank; discharging residue in tank A, and adding Morchella esculenta again; adding water into the extraction tank B to perform third extraction of the extraction tank B, feeding the obtained extract into the extraction tank C to perform second extraction of the extraction tank C, feeding the obtained extract into the extraction tank A to perform first extraction of raw materials in the second tank of the extraction tank A, and feeding the obtained extract into a storage tank; discharging slag from the tank B, adding the morchella raw material again, adding water into the extraction tank C to perform third extraction of the extraction tank C, feeding the obtained extracting solution into the extraction tank A to perform second extraction of the raw material in the second tank of the extraction tank A, feeding the obtained extracting solution serving as an extracting solvent into the extraction tank B to perform first extraction of the extraction tank B, and feeding the obtained extracting solution into a storage tank; discharging slag in a tank C, and adding the morchella raw material again; the extraction is sequentially circulated, and the morchella raw material added into each tank is extracted for three times.
3. The method according to claim 2, wherein the extraction conditions are: carrying out ultrasonic-assisted extraction in the water bath time of the extraction tank at 50-75 ℃ for 0.5-3 h;
the power of the ultrasonic equipment is 200-600 w;
the power of the ultrasonic waves extracted for the first time is 400-600 w;
the power of the second extraction is 300-400 w;
the power of the third extraction is 200-300 w;
the ultrasonic-assisted extraction time is 10-30 min for each extraction;
the total dosage of water in the morchella polysaccharide aqueous extract in the step (1) is calculated according to the ratio of the total dosage g of morchella powder to the total dosage mL of water being 3: 40-50;
in the morchella crude polysaccharide dissolving solution in the step (3), calculating a morchella crude polysaccharide extract and a solvent according to a volume ratio of 1: 4-6;
the protease in the step (4) comprises at least one of trypsin, papain and bromelain;
when the protease is trypsin, adjusting the pH value to 6-8;
when the protease is papain, adjusting the pH value to 4-6;
when the protease is bromelain, adjusting the pH value to 6-8;
the dosage of the trypsin is 10-25U/mg;
the dosage of the papain is 10-25U/mg;
the using amount of the bromelain is 10-25U/mg;
the temperature of the enzymolysis deproteinization in the step (4) is 50-65 ℃;
and (4) the time for enzymolysis and deproteinization in the step (4) is 1-3 h.
4. A morchella polysaccharide extract, which is prepared by the preparation method of any one of claims 1 to 3.
5. The use of the morchella polysaccharide extract of claim 4 in the preparation of a health-care functional preparation for tonifying kidney and strengthening yang.
6. A health functional preparation for invigorating kidney and strengthening yang, comprising an adjuvant and the morchella polysaccharide extract according to claim 4.
7. An effervescent tablet containing morchella polysaccharide extract is characterized by comprising a component A, a sweetening agent and a lubricating agent; the component A comprises the following components in parts by mass: 220-300 parts of morchella esculenta polysaccharide extract, 260-320 parts of citric acid, 160-200 parts of sodium bicarbonate, 180-240 parts of cane sugar and 200-290 parts of isomaltitol; the content of the sweetening agent is 0-1.0% of the mass of the component A, and the content of the lubricating agent is 0-0.7% of the mass of the component A.
8. The effervescent tablet containing morchella polysaccharide extract according to claim 7, wherein the sweetener is sucralose;
the lubricant is magnesium stearate.
9. The effervescent tablet containing the morchella polysaccharide extract according to claim 7, wherein the content of the sweetener is 0.5-1.0% of the component A by mass;
the content of the lubricant is 0.2-0.7% of the component A by mass.
10. The preparation method of the effervescent tablet containing the morchella polysaccharide extract according to any one of claims 7 to 9, characterized by comprising the following steps:
mixing the ground and sieved sweetener, citric acid, cane sugar and isomalt, and adding a wetting agent to prepare wet granules A; mixing morchella polysaccharide and sodium bicarbonate, adding a wetting agent, and preparing wet granules B;
(II) drying and finishing the wet granules A and the wet granules B obtained in the step (I) respectively to obtain two kinds of dry granules;
(III) mixing the two dry particles obtained in step (II) with a lubricant to obtain a total mixture;
(IV) tabletting the total mixture obtained in the step (III) to obtain effervescent tablets containing the morchella polysaccharide extract;
wherein, the wetting agent is 50% (v/v) ethanol solution;
the dosage of the wetting agent is calculated according to the ratio of the wetting agent to the required wetting material being 0.2-0.3 mL: 1 g.
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Application publication date: 20201020 |